基于UPLC-Q-TOF_...困型溃疡性结肠炎大鼠的机制_钟薏文.pdf

中华中医药杂志(原中国医药学报)2023年2月第38卷第2期 CJTCMP,February 2023,Vol.38,No.2 840 8 Blkbas I,Mundorf A,Freund N.Maternal separation in rats induces neurobiological and behavioral changes on the maternal side.Scientific Reports,2020,10(1):2243-22439 Maniam J,Morris M J.Long-term postpartum anxiety and depression-like behavior in mother rats subjected to maternal separation are ameliorated by palatable high fat diet.Behavioural Brain Research,2010,208(1):72-7910 Gur T L,Shay L,Palkar A V,et al.Prenatal stress affects placental cytokines and neurotrophins,commensal microbes,and anxiety-like behavior in adult female offspring.Brain Behav Immun,2017,8(64):50-5811 罗阳,郑乐知,周建伟,等.产后抑郁症与雌激素及单胺递质水平的相关性研究.中华妇产科杂志,2007,42(11):745-74812 胡电,古航,熊英,等.产后抑郁症与孤啡肽及单胺类递质的相关性研究.中国神经精神疾病杂志,2003,29(5):321-32213 符小航,符海鸽,梁亮,等.柴胡加龙骨牡蛎汤加减联合帕罗西汀治疗产后抑郁患者的疗效及对5-羟色胺的影响.现代中西医结合杂志,2021,30(2):188-19114 赵欣媛,张建华,张瑾,等.80例产后抑郁症患者血清雌二醇及5-羟色胺变化分析.国际精神病学杂志,2016,43(5):845-84715 Green A D,Galea L.Adult hippocampal cell proliferation is suppressed with estrogen withdrawal after a hormone-simulated pregnancy.Hormones&Behavior,2008,54(1):203-21116 何娜,魏云,何清林,等.复方乌鳢口服液对产后抑郁症模型大鼠行为学的影响.实用中医药杂志,2020,36(9):1109-111017 曲淼,孙文军,唐启盛.“枢轴运动”理论与产后抑郁症辨治.北京中医药大学学报,2021,44(7):631-63418 杨歆科,唐启盛.论产后抑郁症“补虚祛瘀,颐脑醒神”治法.中华中医药杂志,2012,27(4):1131-113519 王丹,李小黎,赵瑞珍,等.参芪解郁方对产后抑郁症患者雌、孕激素及EPDS评分的影响.北京中医药,2016,35(1):29-3220 曲淼,李净娅,唐启盛,等.Treg/Th17平衡对产后抑郁大鼠的影响及参芪解郁方的细胞免疫调节作用.中华中医药杂志,2015,30(5):1452-145821 侯秀娟,唐启盛,杨歆科,等.参芪解郁方对产后抑郁大鼠单胺递质及其代谢产物变化的干预研究.北京中医药,2013,32(3):182-188（收稿日期：2022年2月23日）基于UPLC-Q-TOF/MS和粪便代谢组学探讨参苓白术散改善脾虚湿困型溃疡性结肠炎大鼠的机制钟薏文1，张文彤1，陈雅茜2，徐霞1，王玮玮3，涂钰1，熊艳玲4，徐国良1，李冰涛1，张启云1，姜丽1（1江西中医药大学中医基础理论分化发展研究中心，江西省中医病因生物学重点实验室，江西省中药 药理学重点实验室，南昌 330004；2南京中医药大学药学院，南京 210000；3井冈山学院 附属医院，吉安 343000；4中南大学湘雅医院临床药理研究所，长沙 410008）摘要：目的：基于超高效液相色谱-四级杆-飞行时间质谱（UPLC-Q-TOF/MS）技术探讨参苓白术散对脾虚湿困型溃疡性结肠炎（SDDR-UC）大鼠粪便中内源性标志物的影响，旨在寻找粪便中差异的生物标志物群，从代谢组学的角度阐述参苓白术散改善SDDR-UC的分子机制。方法：将雄性SD大鼠随机分为5组：正常组，SDDR-UC模型组，参苓白术散低、中、高剂量组。采用病症结合法复制脾虚湿困证模型大鼠，再灌胃5%葡聚糖硫酸钠造成SDDR-UC模型。给药阶段，各治疗组灌胃参苓白术散低、中、高剂量（0.472、0.945、1.89 gkg-1d-1），正常组、模型组则灌胃灭菌纯净水。实验结束前，收集各组大鼠粪便，应用UPLC-Q-TOF/MS方法测定粪便代谢物的含量，结合多维统计分析筛选组间差异代谢物，并进行Metabo Analyst 5.0代谢通路富集分析。结果：共筛选和鉴定出34个潜在生物标志物，主要影响叶酸生物合成、色氨酸代谢、鞘脂代谢、一个叶酸碳库、花生四烯酸代谢5条代谢通路，其中墨蝶呤、6-羟基褪黑素、1-磷酸鞘氨醇、二氢叶酸和12（S）-白三烯B4是与代谢通路密切相关的研究报告通信作者：姜丽，江西省南昌市新建区梅岭大道1688号江西中医药大学中医基础理论分化发展研究中心，邮编：330004电话：0791-87142856，E-mail：内文2.indd 8402023/2/28 10:36:28中华中医药杂志(原中国医药学报)2023年2月第38卷第2期 CJTCMP,February 2023,Vol.38,No.2 841 关键潜在生物标志物。结论：参苓白术散对SDDR-UC的治疗作用可能与其调控炎性因子的释放、抗脂质过氧化和氧化应激损伤、修复受损屏障、维持结肠上皮细胞黏膜的完整性，进而恢复体内正常代谢活动有关。关键词：参苓白术散；脾虚湿困；溃疡性结肠炎；粪便；代谢组学；超高效液相色谱-四级杆-飞行时间质谱基金资助：江西中医药大学校级科技创新团队发展计划（No.CXTD22007），江西省卫生计生委中医药科研课题（No.2021B599），江西省卫生健康委科技计划课题（No.SKJP_220210795）Mechanism of Shenling Baizhu Powder on improving spleen deficiency with dampness retention-ulcerative colitis rats based on UPLC-Q-TOF/MS and fecal metabonomicsZHONG Yi-wen1,ZHANG Wen-tong1,CHEN Ya-xi2,XU Xia1,WANG Wei-wei3,TU Yu1,XIONG Yan-ling4,XU Guo-liang1,LI Bing-tao1,ZHANG Qi-yun1,JIANG Li1(1Research Center for Differentiation and Development of Traditional Chinese Medicine Basic Theory,Key laboratory of TCM Etiopathogenisis,Key Laboratory of Pharmacology of TCM,Jiangxi University of Chinese Medicine,Nanchang 330004,China;2College of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210000,China;3Affiliated Hospital of Jinggangshan University,Jian 343000,China;4Department of Clinical Pharmacology,Xiangya Hospital,Central South University,Changsha 410008,China)Abstract:Objective:Based on Ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS)to investigate the effects of Shenling Baizhu Powder on endogenous markers in feces of spleen deficiency with dampness retention-ulcerative colitis(SDDR-UC)rats,aiming to find different biomarker groups in feces.The molecular mechanism of Shenling Baizhu Powder in improving SDDR-UC was elaborated from the perspective of metabolomics.Methods:Male SD rats were randomly divided into 5 groups:normal group,SDDR-UC model group,low-,medium-and high-dose Shenling Baizhu Powder groups.The SDDR-UC model rats were induced by diet combined with environment intervention and 5%DSS intragastric administration.At the administration stage,the treatment groups were given with low-,medium-,and high-dose(0.472,0.945,1.89 gkg-1d-1)of Shenling Baizhu Powder,while normal group and model group were given sterilized pure water.Before the end of the experiment,the fece of rats were collected,and then detected by UPLC-Q-TOF/MS.The differential metabolites associated with this disease were screened by multi-dimensional statistical analysis.Finally,Metabo Analyst 5.0 was used for metabolic pathway enrichment analysis.Results:In this experiment,34 potential biomarkers were screened and identified,which mainly affected 5 metabolic pathways such as Folate biosynthesis,Tryptophan metabolism,Sphingolipid metabolism,One carbon pool by folate and Arachiidonic acid metabolism.Among the above metabolic pathways,Sepiapterin,6-hydro